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. 1992 Feb;174(4):1172–1178. doi: 10.1128/jb.174.4.1172-1178.1992

Inhibition of the recBCD-dependent activation of Chi recombinational hot spots in SOS-induced cells of Escherichia coli.

R Rinken 1, W Wackernagel 1
PMCID: PMC206409  PMID: 1310498

Abstract

Nucleotide sequences called Chi (5'-GCTGGTGG-3') enhance homologous recombination near their location by the RecBCD enzyme in Escherichia coli (Chi activation). A partial inhibition of Chi activation measured in lambda red gam mutant crosses was observed after treatment of wild-type cells with DNA-damaging agents including UV, mitomycin, and nalidixic acid. Inhibition of Chi activation was not accompanied by an overall decrease of recombination. A lexA3 mutation which blocks induction of the SOS system prevented the inhibition of Chi activation, indicating that an SOS function could be responsible for the inhibition. Overproduction of the RecD subunit of the RecBCD enzyme from a multicopy plasmid carrying the recD gene prevented the induced inhibition of Chi activation, whereas overproduction of RecB or RecC subunits did not. It is proposed that in SOS-induced cells the RecBCD enzyme is modified into a Chi-independent recombination enzyme, with the RecD subunit being the regulatory switch key.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Amundsen S. K., Neiman A. M., Thibodeaux S. M., Smith G. R. Genetic dissection of the biochemical activities of RecBCD enzyme. Genetics. 1990 Sep;126(1):25–40. doi: 10.1093/genetics/126.1.25. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Amundsen S. K., Taylor A. F., Chaudhury A. M., Smith G. R. recD: the gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci U S A. 1986 Aug;83(15):5558–5562. doi: 10.1073/pnas.83.15.5558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Biek D. P., Cohen S. N. Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli. J Bacteriol. 1986 Aug;167(2):594–603. doi: 10.1128/jb.167.2.594-603.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Calsou P., Villaverde A., Defais M. Activated RecA protein may induce expression of a gene that is not controlled by the LexA repressor and whose function is required for mutagenesis and repair of UV-irradiated bacteriophage lambda. J Bacteriol. 1987 Oct;169(10):4816–4821. doi: 10.1128/jb.169.10.4816-4821.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chaudhury A. M., Smith G. R. A new class of Escherichia coli recBC mutants: implications for the role of RecBC enzyme in homologous recombination. Proc Natl Acad Sci U S A. 1984 Dec;81(24):7850–7854. doi: 10.1073/pnas.81.24.7850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cheng K. C., Smith G. R. Cutting of chi-like sequences by the RecBCD enzyme of Escherichia coli. J Mol Biol. 1987 Apr 20;194(4):747–750. doi: 10.1016/0022-2836(87)90252-x. [DOI] [PubMed] [Google Scholar]
  7. Cheng K. C., Smith G. R. Recombinational hotspot activity of Chi-like sequences. J Mol Biol. 1984 Dec 5;180(2):371–377. doi: 10.1016/s0022-2836(84)80009-1. [DOI] [PubMed] [Google Scholar]
  8. Clark A. J. Recombination deficient mutants of E. coli and other bacteria. Annu Rev Genet. 1973;7:67–86. doi: 10.1146/annurev.ge.07.120173.000435. [DOI] [PubMed] [Google Scholar]
  9. Dower N. A., Stahl F. W. Chi activity during transduction-associated recombination. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7033–7037. doi: 10.1073/pnas.78.11.7033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dykstra C. C., Prasher D., Kushner S. R. Physical and biochemical analysis of the cloned recB and recC genes of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):21–27. doi: 10.1128/jb.157.1.21-27.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ginsburg H., Edmiston S. H., Harper J., Mount D. W. Isolation and characterization of an operator-constitutive mutation in the recA gene of E. coli K-12. Mol Gen Genet. 1982;187(1):4–11. doi: 10.1007/BF00384376. [DOI] [PubMed] [Google Scholar]
  12. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  13. Howard-Flanders P., Boyce R. P. DNA repair and genetic recombination: studies on mutants of Escherichia coli defective in these processes. Radiat Res. 1966;(Suppl):156+–156+. [PubMed] [Google Scholar]
  14. Kobayashi I., Murialdo H., Crasemann J. M., Stahl M. M., Stahl F. W. Orientation of cohesive end site cos determines the active orientation of chi sequence in stimulating recA . recBC-mediated recombination in phage lambda lytic infections. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5981–5985. doi: 10.1073/pnas.79.19.5981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kobayashi I., Stahl M. M., Fairfield F. R., Stahl F. W. Coupling with packaging explains apparent nonreciprocality of Chi-stimulated recombination of bacteriophage lambda by RecA and RecBC functions. Genetics. 1984 Dec;108(4):773–794. doi: 10.1093/genetics/108.4.773. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Krueger J. H., Elledge S. J., Walker G. C. Isolation and characterization of Tn5 insertion mutations in the lexA gene of Escherichia coli. J Bacteriol. 1983 Mar;153(3):1368–1378. doi: 10.1128/jb.153.3.1368-1378.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Little J. W., Mount D. W. The SOS regulatory system of Escherichia coli. Cell. 1982 May;29(1):11–22. doi: 10.1016/0092-8674(82)90085-x. [DOI] [PubMed] [Google Scholar]
  18. Lloyd R. G., Buckman C. Overlapping functions of recD, recJ and recN provide evidence of three epistatic groups of genes in Escherichia coli recombination and DNA repair. Biochimie. 1991 Feb-Mar;73(2-3):313–320. doi: 10.1016/0300-9084(91)90218-p. [DOI] [PubMed] [Google Scholar]
  19. Lloyd R. G., Porton M. C., Buckman C. Effect of recF, recJ, recN, recO and ruv mutations on ultraviolet survival and genetic recombination in a recD strain of Escherichia coli K12. Mol Gen Genet. 1988 May;212(2):317–324. doi: 10.1007/BF00334702. [DOI] [PubMed] [Google Scholar]
  20. Lovett S. T., Clark A. J. Genetic analysis of the recJ gene of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):190–196. doi: 10.1128/jb.157.1.190-196.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lovett S. T., Luisi-DeLuca C., Kolodner R. D. The genetic dependence of recombination in recD mutants of Escherichia coli. Genetics. 1988 Sep;120(1):37–45. doi: 10.1093/genetics/120.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lundblad V., Taylor A. F., Smith G. R., Kleckner N. Unusual alleles of recB and recC stimulate excision of inverted repeat transposons Tn10 and Tn5. Proc Natl Acad Sci U S A. 1984 Feb;81(3):824–828. doi: 10.1073/pnas.81.3.824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Malone R. E., Chattoraj D. K., Faulds D. H., Stahl M. M., Stahl F. W. Hotspots for generalized recombination in the Escherichia coli chromosome. J Mol Biol. 1978 Jun 5;121(4):473–491. doi: 10.1016/0022-2836(78)90395-9. [DOI] [PubMed] [Google Scholar]
  24. Markham B. E., Harper J. E., Mount D. W., Sancar G. B., Sancar A., Rupp W. D., Kenyon C. J., Walker G. C. Analysis of mRNA synthesis following induction of the Escherichia coli SOS system. J Mol Biol. 1984 Sep 15;178(2):237–248. doi: 10.1016/0022-2836(84)90142-6. [DOI] [PubMed] [Google Scholar]
  25. Mount D. W. A mutant of Escherichia coli showing constitutive expression of the lysogenic induction and error-prone DNA repair pathways. Proc Natl Acad Sci U S A. 1977 Jan;74(1):300–304. doi: 10.1073/pnas.74.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mount D. W., Low K. B., Edmiston S. J. Dominant mutations (lex) in Escherichia coli K-12 which affect radiation sensitivity and frequency of ultraviolet lght-induced mutations. J Bacteriol. 1972 Nov;112(2):886–893. doi: 10.1128/jb.112.2.886-893.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Palas K. M., Kushner S. R. Biochemical and physical characterization of exonuclease V from Escherichia coli. Comparison of the catalytic activities of the RecBC and RecBCD enzymes. J Biol Chem. 1990 Feb 25;265(6):3447–3454. [PubMed] [Google Scholar]
  28. Ponticelli A. S., Schultz D. W., Taylor A. F., Smith G. R. Chi-dependent DNA strand cleavage by RecBC enzyme. Cell. 1985 May;41(1):145–151. doi: 10.1016/0092-8674(85)90069-8. [DOI] [PubMed] [Google Scholar]
  29. Quiñones A., Jüterbock W. R., Messer W. Expression of the dnaA gene of Escherichia coli is inducible by DNA damage. Mol Gen Genet. 1991 May;227(1):9–16. doi: 10.1007/BF00260699. [DOI] [PubMed] [Google Scholar]
  30. Rinken R., de Vries J., Weichenhan D., Wackernagel W. The recA-recBCD dependent recombination pathways of Serratia marcescens and Proteus mirabilis in Escherichia coli: functions of hybrid enzymes and hybrid pathways. Biochimie. 1991 Apr;73(4):375–384. doi: 10.1016/0300-9084(91)90104-9. [DOI] [PubMed] [Google Scholar]
  31. Schultz D. W., Taylor A. F., Smith G. R. Escherichia coli RecBC pseudorevertants lacking chi recombinational hotspot activity. J Bacteriol. 1983 Aug;155(2):664–680. doi: 10.1128/jb.155.2.664-680.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smith G. R. Mechanism and control of homologous recombination in Escherichia coli. Annu Rev Genet. 1987;21:179–201. doi: 10.1146/annurev.ge.21.120187.001143. [DOI] [PubMed] [Google Scholar]
  33. Stahl F. W. Special sites in generalized recombination. Annu Rev Genet. 1979;13:7–24. doi: 10.1146/annurev.ge.13.120179.000255. [DOI] [PubMed] [Google Scholar]
  34. Stahl F. W., Stahl M. M., Malone R. E., Crasemann J. M. Directionality and nonreciprocality of Chi-stimulated recombination in phage lambda. Genetics. 1980 Feb;94(2):235–248. doi: 10.1093/genetics/94.2.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Stahl F. W., Stahl M. M. Recombination pathway specificity of Chi. Genetics. 1977 Aug;86(4):715–725. doi: 10.1093/genetics/86.4.715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stahl F. W., Thomason L. C., Siddiqi I., Stahl M. M. Further tests of a recombination model in which chi removes the RecD subunit from the RecBCD enzyme of Escherichia coli. Genetics. 1990 Nov;126(3):519–533. doi: 10.1093/genetics/126.3.519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Taylor A. F., Schultz D. W., Ponticelli A. S., Smith G. R. RecBC enzyme nicking at Chi sites during DNA unwinding: location and orientation-dependence of the cutting. Cell. 1985 May;41(1):153–163. doi: 10.1016/0092-8674(85)90070-4. [DOI] [PubMed] [Google Scholar]
  38. Thaler D. S., Sampson E., Siddiqi I., Rosenberg S. M., Thomason L. C., Stahl F. W., Stahl M. M. Recombination of bacteriophage lambda in recD mutants of Escherichia coli. Genome. 1989;31(1):53–67. doi: 10.1139/g89-013. [DOI] [PubMed] [Google Scholar]
  39. Thaler D. S., Stahl F. W. DNA double-chain breaks in recombination of phage lambda and of yeast. Annu Rev Genet. 1988;22:169–197. doi: 10.1146/annurev.ge.22.120188.001125. [DOI] [PubMed] [Google Scholar]
  40. Thoms B., Wackernagel W. Regulatory role of recF in the SOS response of Escherichia coli: impaired induction of SOS genes by UV irradiation and nalidixic acid in a recF mutant. J Bacteriol. 1987 Apr;169(4):1731–1736. doi: 10.1128/jb.169.4.1731-1736.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Thoms B., Wackernagel W. UV-induced allevation of lambda restriction in Escherichia coli K-12: kinetics of induction and specificity of this SOS function. Mol Gen Genet. 1982;186(1):111–117. doi: 10.1007/BF00422921. [DOI] [PubMed] [Google Scholar]
  42. Walker G. C. Inducible DNA repair systems. Annu Rev Biochem. 1985;54:425–457. doi: 10.1146/annurev.bi.54.070185.002233. [DOI] [PubMed] [Google Scholar]
  43. Walker G. C. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. doi: 10.1128/mr.48.1.60-93.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Weichenhan D., Wackernagel W. Cloning of the recB, recC, and recD genes from Proteus mirabilis in Escherichia coli: in vivo formation of active hybrid enzymes. J Bacteriol. 1988 Mar;170(3):1412–1414. doi: 10.1128/jb.170.3.1412-1414.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Witkin E. M. Ultraviolet mutagenesis and inducible DNA repair in Escherichia coli. Bacteriol Rev. 1976 Dec;40(4):869–907. doi: 10.1128/br.40.4.869-907.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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